Medium, for culturing corneal endothelial cells, containing conditioned medium from mesenchymal stem cells

Abstract

[Problem] To provide, in order to manufacture cells to transplant into patients with corneal endothelial failure, a medium used to culture corneal endothelial cells obtained from human corneal tissue and grow said cells while maintaining the morphology thereof as corneal endothelial cells. [Solution] A medium containing a conditioned medium from mesenchymal stem cells; and a method in which said medium is used to culture corneal endothelial cells.

The invention claimed is: 
     
       1. A method for culturing human corneal endothelial cells, which comprises
 a step for preparing a conditioned medium by culturing a mesenchymal stem cell in a medium containing chondroitin sulfate, 
 a step for separating the human corneal endothelial cells from a corneal tissue, and 
 a step for propagating the separated human corneal endothelial cells by a culture using a medium for corneal endothelial cell culture consisting of the conditioned medium of mesenchymal stem cells or containing the conditioned medium wherein the mesenchymal stem cell is a human mesenchymal stem cell and wherein the concentration of chondroitin sulfate is 0.04 to 0.12 % (w/v). 
 
     
     
       2. The method according to  claim 1 , wherein the medium for corneal endothelial cell culture contains the conditioned medium in a proportion of 0.5 to 50 % (v/v). 
     
     
       3. The method according to  claim 1 , wherein the medium for corneal endothelial cell culture contains the conditioned medium in a proportion of 1to 10 % (v/v). 
     
     
       4. The method according to  claim 1 , wherein the concentration of chondroitin sulfate is 0.08 % (w/v). 
     
     
       5. The method according to  claim 1 , wherein the mesenchymal stem cell is a mesenchymal stem cell derived from human bone marrow. 
     
     
       6. A method for culturing human corneal endothelial cells, which comprises
 a step for preparing a conditioned medium by culturing a mesenchymal stem cell in a medium containing chondroitin sulfate, 
 a step for preparing a concentrated solution by filtration concentration of the conditioned medium, 
 a step for separating the human corneal endothelial cells from a corneal tissue, and 
 a step for propagating the separated human corneal endothelial cells by a culture using a medium for corneal endothelial cell culture containing the concentrated solution wherein the mesenchymal stem cell is a human mesenchymal stem cell and wherein the concentration of chondroitin sulfate is 0.04 to 0.12 % (w/v). 
 
     
     
       7. The method according to  claim 6 , wherein the concentration rate in the filtration concentration is 2 to 30 times. 
     
     
       8. The method according to  claim 6 , wherein the concentration rate in the filtration concentration is 15 to 20 times. 
     
     
       9. The method according to  claim 6 , wherein the medium for corneal endothelial cell culture contains the concentrated solution at a proportion of 0.5 to 50 % (v/v). 
     
     
       10. The method according to  claim 6 , wherein the medium for corneal endothelial cell culture contains the concentrated solution at a proportion of 1 to 10 % (v/v). 
     
     
       11. The method according to  claim 6 , wherein the concentration of chondroitin sulfate is 0.08 % (w/v). 
     
     
       12. The method according to  claim 6 , wherein the mesenchymal stem cell is a mesenchymal stem cell derived from human bone marrow. 
     
     
       13. The method according to  claim 1 , wherein the medium for corneal endothelial cell culture is not added with nerve growth factor. 
     
     
       14. The method according to  claim 6 , wherein the medium for corneal endothelial cell culture is not added with nerve growth factor.